The rapid range expansion of West Nile Virus has raised interest in understanding the population dynamics and dispersal patterns of emerging infectious diseases by wildlife. We analyzed different ecological and evolutionary factors related to West Nile Virus neutralizing antibody prevalence in 72 bird species sampled in southern Spain. Prevalence of antibodies reached its maximum during the autumn and winter in comparison to summer months. Prevalence of antibodies was directly related to body mass and migratory behaviour. The greater prevalence of antibodies observed in summer migrants can be explained, among other factors, by the diversity of localities involved in their life cycles or the geographic areas visited during their migrations. Greater prevalence in larger species was explained by their longevity because the relationship was already significant when analyzing only first year birds, and probably also involved a high attraction to vectors by larger hosts. Coloniality and winter gregarism were unrelated to the prevalence of antibodies against this highly host generalist pathogen. Evolutionary relationships between species were unrelated to differences in the prevalence of antibodies. Our results suggest larger species as good candidates for easy, faster and cheaper monitoring of local, seasonal and annual changes in WN virus serology.